QIAamp DSP Virus Kit

It is possible to use a general laboratory vacuum system with the QIAamp DSP Virus Kit. However, this will require validation in-house. The CE-marked QIAamp Kits have been developed, field-tested and validated with the QIAvac 24 Plus vacuum system (QIAvac 24 Plus, QIAvac Connecting System and Vacuum Pump). Using this system with the QIAamp DSP Virus Kit will avoid lengthy validation procedures for the enduser.

When processing samples with the QIAamp DSP DNA Blood Mini Kit and the QIAamp DSP Virus Kit, the vacuum pressure should be below -800 mbar. The QIAvac 24 Plus and the QIAvac Connecting System should be tested according to Appendix B of the QIAvac 24 Plus Handbook before performing a nucleic acid purification procedure.

If you would like to use an equivalent general laboratory vacuum system, make sure that the minimum vacuum pressure is reached. Performance of the QIAamp DSP DNA Blood Mini or Virus Kits in combination with a house-vacuum system has to be validated by the researcher.

QIAGEN Proteinase K is a subtilisin-type protease, which cleaves at the carboxyl side of hydrophobic, aliphatic and aromatic amino acids. It is particularly suitable for short digestion times. It possesses a high specific activity over a wide range of temperatures and pH values with substantially increased activity at higher temperature. Soluble calcium is not essential for enzymatic activity. This means that EDTA, which is used to inhibit Mg2+-dependent enzymes such as nucleases, will not inhibit Proteinase K activity.

QIAGEN Protease is a broad-specificity Serine protease with high activity, cleaving preferentially at neutral and acidic residues. It is an economical alternative to Proteinase K for isolation of native DNA and RNA from a variety of samples.

For more information on activity of QIAGEN Protease and Proteinase K in various buffers please click here.

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

For RNA isolated on the BioRobot EZ1 and BioRobot M48:

The RNA can be directly applied to the Agilent Bioanalyzer, since it is being denatured in the final protocol steps of these isolation procedures.

For RNA prepared with all other QIAGEN RNA Isolation Products:

We recommend to denature the samples in a water bath for 2 min at 70°C, and then place them directly on ice prior to loading them onto the Agilent Bioanalyzer.